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Use for dendropanax morbifera extract for adjusting 15- hydroxyprostaglandin dehydrogenase and pge 2 activity

Active Publication Date: 2015-07-23
INDUSRTY ACADEMIC COOPERATION FOUND CHOSUN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is for a Dendropanax morbifera extract that can increase the levels of PGE2 within cells and can also inhibit the activity of an enzyme called 15-PGDH that breaks down PGE2. This results in a reduction of PGE2 levels. The extract can therefore be used to treat and prevent various diseases caused by increased levels of PGE2 such as wounds, burns, oral ulcers, peptic ulcers, and gastritis. The technical effect of this invention is the discovery of a new way to modulate the levels of PGE2 and 15-PGDH, which could have important therapeutic benefits in several areas of medicine.

Problems solved by technology

To date, however, neither the molecular mechanisms of a Dendropanax morbifera extract involved in regulating the activity of 15-PGDH and PGE2 were identified nor were disease treatments based on such molecular mechanisms reported.

Method used

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  • Use for dendropanax morbifera extract for adjusting 15- hydroxyprostaglandin dehydrogenase and pge 2 activity
  • Use for dendropanax morbifera extract for adjusting 15- hydroxyprostaglandin dehydrogenase and pge 2 activity
  • Use for dendropanax morbifera extract for adjusting 15- hydroxyprostaglandin dehydrogenase and pge 2 activity

Examples

Experimental program
Comparison scheme
Effect test

example 1

Preparation of Dendropanax morbifera Leaf Extract

[0115](1) Preparation of Extract

[0116]Collected D. morbifera leaves were dried in the shade at room temperature. Dried leaves were extracted with methanol for 3 times. The methanol extracts described above were filtered through Whatman No. 1 filter paper, and combined methanol extracts above described were concentrated in vacuum using rotary evaporator.

[0117]Methanol extracts were suspended in water and then fractionated according to the polarity by hexane, diethyl ether, ethyl acetate, n-butanol and water. Each of the extracts was concentrated using a rotary evaporator. This process is illustrated in FIG. 4.

[0118](2) Fractionation and Separation Process

[0119]Using gradient solvent system [CHCl3 (0.1% HCOOH) 100% through CHCl3 (0.1% HCOOH)-MeOH (0.1% HCOOH) / 55:45 for 120 min], 10 fractions were provided (DEE01-DEE10) by conducting the activity-guided fractionation under 254 and 280 nm with Isolera Flash Purification system (BIOTAGE, S...

example 2

Identification of 15-PGDH Expression

[0124](1) Expression and Purification of 15-PGDH

[0125]Escherichia coli BL-21 DE3 cells were transformed by a PGEX-2T expression vector including recombinant 15-PGDH between BamH I and EcoR I as illustrated in FIG. 5.

[0126]The cells were grown in 500 mL medium containing 50 μg / mL ampicillin at 37° C. and 220 rpm until the O.D. at 600 nm reached 0.6. IPTG (isoprophyl β-D-thiogalactoside, 1M stock solution) was added and the cells were grown for 12 hours at 25° C.

[0127]Later, the cells were centrifuged at 4000×g, 4° C. for 30 minutes and collected and the cell pellet was resuspended in 20 mL cold lysis buffer (1× PBS buffer pH 7.4 containing 1 mM EDTA and 0.1 mM DTT) and disrupted by ultrasonication.

[0128]The supernatant was slowly applied to glutathione-sepharose 4B column and equilibrated at 4° C. using lysis buffer. The column described above was rinsed with lysis buffer until O.D. at 280 nm was measured lower than 0.005. Then, 15-PGDH was eluted ...

example 3

Analysis of Cell Viability

[0137](1) Cell Culture

[0138]The HaCaT cells, human keratinocyte cell line, were cultured using DMEM (Dulbecco's modified Eagle's media) in 5% CO2 humidified incubator at 37° C.

[0139]The A549 cells originated from adenocarcinoma-like human alveolar type II were cultured in RPMI medium, and 5% CO2 humidified incubator at 37° C. And then, LNCaP•FGC (androgen-dependent prostate cancer) cells were cultured in RPMI medium and 5% CO2 humidified incubator at 37° C. All culture media were supplemented with heat-inactivated FBS and 100 μg / mL penicillin

[0140](2) Cytotoxicity Test

[0141]The cytotoxicity was determined by MTT assay (Mosmann, 1983).

[0142]The HaCaT cells (1×104 / mL) and LNCaP•FGC cells (4×104 / mL) were seeded per 96 μL of DMEM medium on 96-well plate. After overnight culture, drugs were treated for 72 hours and cultured in 10 μL of MTT (5 mg / mL stock solution) for 4 hours. Then, the medium was removed and formazan was dissolved by adding 150 μL of DMSO. Usin...

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Abstract

Provided is a 15-PGDH activity adjusting function of a Dendropanax morbifera extract. More specifically, since it has been confirmed that the Dendropanax morbifera extract has an outstanding effect in suppressing 15-PGDH activity and so increasing PGE2, a composition of the present invention which contains the Dendropanax morbifera extract as an active ingredient has the advantage of being able to be used for producing a functional health food and a therapeutic agent for a disease associated with PGE2.

Description

TECHNICAL FIELD[0001]The present invention relates to a function of a Dendropanax morbifera extract, by which the activities of 15-hydroxyprostaglandin dehydrogenase (15-PGDH) and PGE2 are regulated, and more specifically, it relates to a composition including a Dendropanax morbifera extract as an active ingredient, in which the Dendropanax morbifera extract allows the activity of 15-PGDH to be inhibited to increase the level of PGE2, thereby preventing and treating the diseases associated with PGE2, or an use thereof.BACKGROUND ART[0002]Dendropanax morbifera Lev., which is a member of a Korea angelica tree, is an evergreen broad-leaved tree that never shed its leaves even during the winter, and grows naturally in the southern coastal area and Jeju-do, Korea, and when the bark of a tree is wounded, a yellow liquid resin is released, and called as Hwangchil ( a yellow dye).[0003]Hwangchil has been used as a precious paint to give off a golden color for armor, helmet, and other metal ...

Claims

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Application Information

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IPC IPC(8): A61K36/25A23L1/30
CPCA61K36/25A23V2002/00A23L1/3002A23L33/105A61P17/02A61P29/00A23V2200/30A23V2200/3202A23V2250/21
Inventor CHOI, CHEOL-HEECHO, HOONMOON, YOUNG-SOOKHAN, JI-WONHAN, JU-HEE
Owner INDUSRTY ACADEMIC COOPERATION FOUND CHOSUN UNIV
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